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. 1997 Nov 3;16(21):6394–6406. doi: 10.1093/emboj/16.21.6394

The chaperone-assisted membrane release and folding pathway is sensed by two signal transduction systems.

C H Jones 1, P N Danese 1, J S Pinkner 1, T J Silhavy 1, S J Hultgren 1
PMCID: PMC1170246  PMID: 9351822

Abstract

The assembly of interactive protein subunits into extracellular structures, such as pilus fibers in the Enterobacteriaceae, is dependent on the activity of PapD-like periplasmic chaperones. The ability of PapD to undergo a beta zippering interaction with the hydrophobic C-terminus of pilus subunits facilitates their folding and release from the cytoplasmic membrane into the periplasm. In the absence of the chaperone, subunits remained tethered to the membrane and were driven off-pathway via non-productive interactions. These off-pathway reactions were detrimental to cell growth; wild-type growth was restored by co-expression of PapD. Subunit misfolding in the absence of PapD was sensed by two parallel pathways: the Cpx two-component signaling system and the sigma E modulatory pathway.

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Selected References

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  1. Bullitt E., Jones C. H., Striker R., Soto G., Jacob-Dubuisson F., Pinkner J., Wick M. J., Makowski L., Hultgren S. J. Development of pilus organelle subassemblies in vitro depends on chaperone uncapping of a beta zipper. Proc Natl Acad Sci U S A. 1996 Nov 12;93(23):12890–12895. doi: 10.1073/pnas.93.23.12890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cosma C. L., Danese P. N., Carlson J. H., Silhavy T. J., Snyder W. B. Mutational activation of the Cpx signal transduction pathway of Escherichia coli suppresses the toxicity conferred by certain envelope-associated stresses. Mol Microbiol. 1995 Nov;18(3):491–505. doi: 10.1111/j.1365-2958.1995.mmi_18030491.x. [DOI] [PubMed] [Google Scholar]
  3. Danese P. N., Silhavy T. J. The sigma(E) and the Cpx signal transduction systems control the synthesis of periplasmic protein-folding enzymes in Escherichia coli. Genes Dev. 1997 May 1;11(9):1183–1193. doi: 10.1101/gad.11.9.1183. [DOI] [PubMed] [Google Scholar]
  4. Danese P. N., Snyder W. B., Cosma C. L., Davis L. J., Silhavy T. J. The Cpx two-component signal transduction pathway of Escherichia coli regulates transcription of the gene specifying the stress-inducible periplasmic protease, DegP. Genes Dev. 1995 Feb 15;9(4):387–398. doi: 10.1101/gad.9.4.387. [DOI] [PubMed] [Google Scholar]
  5. De Las Peñas A., Connolly L., Gross C. A. The sigmaE-mediated response to extracytoplasmic stress in Escherichia coli is transduced by RseA and RseB, two negative regulators of sigmaE. Mol Microbiol. 1997 Apr;24(2):373–385. doi: 10.1046/j.1365-2958.1997.3611718.x. [DOI] [PubMed] [Google Scholar]
  6. Dodd D. C., Bassford P. J., Jr, Eisenstein B. I. Dependence of secretion and assembly of type 1 fimbrial subunits of Escherichia coli on normal protein export. J Bacteriol. 1984 Sep;159(3):1077–1079. doi: 10.1128/jb.159.3.1077-1079.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dodson K. W., Jacob-Dubuisson F., Striker R. T., Hultgren S. J. Outer-membrane PapC molecular usher discriminately recognizes periplasmic chaperone-pilus subunit complexes. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3670–3674. doi: 10.1073/pnas.90.8.3670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fürste J. P., Pansegrau W., Frank R., Blöcker H., Scholz P., Bagdasarian M., Lanka E. Molecular cloning of the plasmid RP4 primase region in a multi-host-range tacP expression vector. Gene. 1986;48(1):119–131. doi: 10.1016/0378-1119(86)90358-6. [DOI] [PubMed] [Google Scholar]
  9. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  10. Hultgren S. J., Abraham S., Caparon M., Falk P., St Geme J. W., 3rd, Normark S. Pilus and nonpilus bacterial adhesins: assembly and function in cell recognition. Cell. 1993 Jun 4;73(5):887–901. doi: 10.1016/0092-8674(93)90269-v. [DOI] [PubMed] [Google Scholar]
  11. Hultgren S. J., Lindberg F., Magnusson G., Kihlberg J., Tennent J. M., Normark S. The PapG adhesin of uropathogenic Escherichia coli contains separate regions for receptor binding and for the incorporation into the pilus. Proc Natl Acad Sci U S A. 1989 Jun;86(12):4357–4361. doi: 10.1073/pnas.86.12.4357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hultgren S. J., Normark S., Abraham S. N. Chaperone-assisted assembly and molecular architecture of adhesive pili. Annu Rev Microbiol. 1991;45:383–415. doi: 10.1146/annurev.mi.45.100191.002123. [DOI] [PubMed] [Google Scholar]
  13. Hung D. L., Knight S. D., Woods R. M., Pinkner J. S., Hultgren S. J. Molecular basis of two subfamilies of immunoglobulin-like chaperones. EMBO J. 1996 Aug 1;15(15):3792–3805. [PMC free article] [PubMed] [Google Scholar]
  14. Jacob-Dubuisson F., Heuser J., Dodson K., Normark S., Hultgren S. Initiation of assembly and association of the structural elements of a bacterial pilus depend on two specialized tip proteins. EMBO J. 1993 Mar;12(3):837–847. doi: 10.1002/j.1460-2075.1993.tb05724.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jacob-Dubuisson F., Pinkner J., Xu Z., Striker R., Padmanhaban A., Hultgren S. J. PapD chaperone function in pilus biogenesis depends on oxidant and chaperone-like activities of DsbA. Proc Natl Acad Sci U S A. 1994 Nov 22;91(24):11552–11556. doi: 10.1073/pnas.91.24.11552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Jones C. H., Pinkner J. S., Nicholes A. V., Slonim L. N., Abraham S. N., Hultgren S. J. FimC is a periplasmic PapD-like chaperone that directs assembly of type 1 pili in bacteria. Proc Natl Acad Sci U S A. 1993 Sep 15;90(18):8397–8401. doi: 10.1073/pnas.90.18.8397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Kolmar H., Waller P. R., Sauer R. T. The DegP and DegQ periplasmic endoproteases of Escherichia coli: specificity for cleavage sites and substrate conformation. J Bacteriol. 1996 Oct;178(20):5925–5929. doi: 10.1128/jb.178.20.5925-5929.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kuehn M. J., Heuser J., Normark S., Hultgren S. J. P pili in uropathogenic E. coli are composite fibres with distinct fibrillar adhesive tips. Nature. 1992 Mar 19;356(6366):252–255. doi: 10.1038/356252a0. [DOI] [PubMed] [Google Scholar]
  19. Kuehn M. J., Normark S., Hultgren S. J. Immunoglobulin-like PapD chaperone caps and uncaps interactive surfaces of nascently translocated pilus subunits. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10586–10590. doi: 10.1073/pnas.88.23.10586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kuehn M. J., Ogg D. J., Kihlberg J., Slonim L. N., Flemmer K., Bergfors T., Hultgren S. J. Structural basis of pilus subunit recognition by the PapD chaperone. Science. 1993 Nov 19;262(5137):1234–1241. doi: 10.1126/science.7901913. [DOI] [PubMed] [Google Scholar]
  21. Kuroiwa T., Sakaguchi M., Mihara K., Omura T. Systematic analysis of stop-transfer sequence for microsomal membrane. J Biol Chem. 1991 May 15;266(14):9251–9255. [PubMed] [Google Scholar]
  22. Lindberg F., Tennent J. M., Hultgren S. J., Lund B., Normark S. PapD, a periplasmic transport protein in P-pilus biogenesis. J Bacteriol. 1989 Nov;171(11):6052–6058. doi: 10.1128/jb.171.11.6052-6058.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lipinska B., Zylicz M., Georgopoulos C. The HtrA (DegP) protein, essential for Escherichia coli survival at high temperatures, is an endopeptidase. J Bacteriol. 1990 Apr;172(4):1791–1797. doi: 10.1128/jb.172.4.1791-1797.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Matsuyama S., Tajima T., Tokuda H. A novel periplasmic carrier protein involved in the sorting and transport of Escherichia coli lipoproteins destined for the outer membrane. EMBO J. 1995 Jul 17;14(14):3365–3372. doi: 10.1002/j.1460-2075.1995.tb07342.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mecsas J., Rouviere P. E., Erickson J. W., Donohue T. J., Gross C. A. The activity of sigma E, an Escherichia coli heat-inducible sigma-factor, is modulated by expression of outer membrane proteins. Genes Dev. 1993 Dec;7(12B):2618–2628. doi: 10.1101/gad.7.12b.2618. [DOI] [PubMed] [Google Scholar]
  26. Missiakas D., Betton J. M., Raina S. New components of protein folding in extracytoplasmic compartments of Escherichia coli SurA, FkpA and Skp/OmpH. Mol Microbiol. 1996 Aug;21(4):871–884. doi: 10.1046/j.1365-2958.1996.561412.x. [DOI] [PubMed] [Google Scholar]
  27. Missiakas D., Mayer M. P., Lemaire M., Georgopoulos C., Raina S. Modulation of the Escherichia coli sigmaE (RpoE) heat-shock transcription-factor activity by the RseA, RseB and RseC proteins. Mol Microbiol. 1997 Apr;24(2):355–371. doi: 10.1046/j.1365-2958.1997.3601713.x. [DOI] [PubMed] [Google Scholar]
  28. Missiakas D., Raina S. Signal transduction pathways in response to protein misfolding in the extracytoplasmic compartments of E. coli: role of two new phosphoprotein phosphatases PrpA and PrpB. EMBO J. 1997 Apr 1;16(7):1670–1685. doi: 10.1093/emboj/16.7.1670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Pogliano J., Lynch A. S., Belin D., Lin E. C., Beckwith J. Regulation of Escherichia coli cell envelope proteins involved in protein folding and degradation by the Cpx two-component system. Genes Dev. 1997 May 1;11(9):1169–1182. doi: 10.1101/gad.11.9.1169. [DOI] [PubMed] [Google Scholar]
  30. Pugsley A. P., Possot O. The general secretory pathway of Klebsiella oxytoca: no evidence for relocalization or assembly of pilin-like PulG protein into a multiprotein complex. Mol Microbiol. 1993 Nov;10(3):665–674. doi: 10.1111/j.1365-2958.1993.tb00938.x. [DOI] [PubMed] [Google Scholar]
  31. Raina S., Missiakas D., Baird L., Kumar S., Georgopoulos C. Identification and transcriptional analysis of the Escherichia coli htrE operon which is homologous to pap and related pilin operons. J Bacteriol. 1993 Aug;175(16):5009–5021. doi: 10.1128/jb.175.16.5009-5021.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Raina S., Missiakas D., Georgopoulos C. The rpoE gene encoding the sigma E (sigma 24) heat shock sigma factor of Escherichia coli. EMBO J. 1995 Mar 1;14(5):1043–1055. doi: 10.1002/j.1460-2075.1995.tb07085.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Roberts J. A., Marklund B. I., Ilver D., Haslam D., Kaack M. B., Baskin G., Louis M., Möllby R., Winberg J., Normark S. The Gal(alpha 1-4)Gal-specific tip adhesin of Escherichia coli P-fimbriae is needed for pyelonephritis to occur in the normal urinary tract. Proc Natl Acad Sci U S A. 1994 Dec 6;91(25):11889–11893. doi: 10.1073/pnas.91.25.11889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rouvière P. E., De Las Peñas A., Mecsas J., Lu C. Z., Rudd K. E., Gross C. A. rpoE, the gene encoding the second heat-shock sigma factor, sigma E, in Escherichia coli. EMBO J. 1995 Mar 1;14(5):1032–1042. doi: 10.1002/j.1460-2075.1995.tb07084.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Slauch J. M., Silhavy T. J. cis-acting ompF mutations that result in OmpR-dependent constitutive expression. J Bacteriol. 1991 Jul;173(13):4039–4048. doi: 10.1128/jb.173.13.4039-4048.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Slonim L. N., Pinkner J. S., Brändén C. I., Hultgren S. J. Interactive surface in the PapD chaperone cleft is conserved in pilus chaperone superfamily and essential in subunit recognition and assembly. EMBO J. 1992 Dec;11(13):4747–4756. doi: 10.1002/j.1460-2075.1992.tb05580.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Snyder W. B., Davis L. J., Danese P. N., Cosma C. L., Silhavy T. J. Overproduction of NlpE, a new outer membrane lipoprotein, suppresses the toxicity of periplasmic LacZ by activation of the Cpx signal transduction pathway. J Bacteriol. 1995 Aug;177(15):4216–4223. doi: 10.1128/jb.177.15.4216-4223.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Snyder W. B., Silhavy T. J. Enhanced export of beta-galactosidase fusion proteins in prlF mutants is Lon dependent. J Bacteriol. 1992 Sep;174(17):5661–5668. doi: 10.1128/jb.174.17.5661-5668.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Stader J., Benson S. A., Silhavy T. J. Kinetic analysis of lamB mutants suggests the signal sequence plays multiple roles in protein export. J Biol Chem. 1986 Nov 15;261(32):15075–15080. [PubMed] [Google Scholar]
  40. Strauch K. L., Beckwith J. An Escherichia coli mutation preventing degradation of abnormal periplasmic proteins. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1576–1580. doi: 10.1073/pnas.85.5.1576. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Strauch K. L., Johnson K., Beckwith J. Characterization of degP, a gene required for proteolysis in the cell envelope and essential for growth of Escherichia coli at high temperature. J Bacteriol. 1989 May;171(5):2689–2696. doi: 10.1128/jb.171.5.2689-2696.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Weis W., Brown J. H., Cusack S., Paulson J. C., Skehel J. J., Wiley D. C. Structure of the influenza virus haemagglutinin complexed with its receptor, sialic acid. Nature. 1988 Jun 2;333(6172):426–431. doi: 10.1038/333426a0. [DOI] [PubMed] [Google Scholar]
  43. Witholt B., Boekhout M., Brock M., Kingma J., Heerikhuizen H. V., Leij L. D. An efficient and reproducible procedure for the formation of spheroplasts from variously grown Escherichia coli. Anal Biochem. 1976 Jul;74(1):160–170. doi: 10.1016/0003-2697(76)90320-1. [DOI] [PubMed] [Google Scholar]
  44. Xu Z., Jones C. H., Haslam D., Pinkner J. S., Dodson K., Kihlberg J., Hultgren S. J. Molecular dissection of PapD interaction with PapG reveals two chaperone-binding sites. Mol Microbiol. 1995 Jun;16(5):1011–1020. doi: 10.1111/j.1365-2958.1995.tb02326.x. [DOI] [PubMed] [Google Scholar]
  45. Zoller M. J., Smith M. Oligonucleotide-directed mutagenesis of DNA fragments cloned into M13 vectors. Methods Enzymol. 1983;100:468–500. doi: 10.1016/0076-6879(83)00074-9. [DOI] [PubMed] [Google Scholar]

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